Comparison of two anti-T-2 toxin immunoglobulins by direct ELISA

Summary Monoclonal anti-T-2 IgGs produced from 12C12 and 15H6 hybridomas were compared by enzyme-linked immunosorbent assay. Binding activity was linear from 0.005 to 0.25 g protein/ml with 12C12 and from 0.005 to 0.09 g protein/ml with 15H6. The quantity of T-2 toxin (g protein/ml) required for one-half maximum binding activity of 15H6 (0.0875 g protein/ml) was approximately 68% that of 12C12 (g protein/ml).     

Effect of thidiazuron on vegetative tissue-derived somatic embryogenesis and flowering of bamboo Bambusa edulis

Current research on somatic embryogenesis of bamboo uses reproductive tissue as explants. However, it was hard to obtain the explant. Shoots of a local accession (3–4 m high) were used for multiple shoot production. In order to obtain embryogenic callus, nodal and internodal tissues from in vitro plantlets were placed on Murashige and Skoog (MS) medium supplemented with 9.2 M kinetin (KN), 13.6 M 2,4-dichlorophenoxyacetic acid (2,4-D), 0.1% (v/v) coconut milk, and 6% (w/v) sucrose. We studied the effects of sucrose and thidiazuron (TDZ) on callus proliferation. Optimal additives to the MS medium for embryogenic callus proliferation were 0.046 M TDZ, 13.6 M 2,4-D and 3% (w/v) sucrose. TDZ also promoted the germination of bamboo somatic embryos. The germination rate of the somatic embryos exceeded 80% on MS-based medium supplemented with 0.455M TDZ. Naphthaleneacetic acid (NAA) reduced germination. Well-developed plantlets were successfully transferred to soil. There was no albino mutant in subsequent culture. In vitro regenerants and potted plants flowered, but no seeds were produced.     

Callus induction and plant regeneration in Vetiveria zizanioides

Callus induction was obtained from basal parts of Vetiveria zizanioides Stapf. leaves cultured on Murashige and Skoog (MS) medium supplemented with 9.0 M 2,4-dichlorophenoxyacetic acid (2,4-d), 5.7 M indoleacetic acid (IAA) and 4.6 M kinetin. Calli were maintained on MS medium with the addition of 0.9 M 2,4-d and 2.3 M kinetin. Shoot formation was obtained from fast growing 14-day-old callus on the same basal medium supplemented with 0.9 M 2,4-d and 9.3 M kinetin. Embryo-like structures were observed. When transferred to basal medium, shoots readily developed roots. Fully developed regenerated plants were then successfully established in soil.     

Mitochondria in fine afferent nerve fibres of the knee joint in the cat: a quantitative electron-microscopical examination

The distribution of mitochondria, their content and concentration (expressed as the ratio of the mean volume of mitochondria and the surface of the sensory axon) were determined in group-III and-IV nerve fibres innervating the knee joint capsule in the cat. Mitochondria mainly accumulated in axonal swellings (beads) and end bulbs of the terminal branches. Between single nerve fibres, marked differences in the content and the concentration of mitochondria were obtained in proximal portions (inside of the perineurium) and in distal portions (unmyelinated sensory endings). In group-III nerve fibres, the mitochondrial concentration ranged from 0.005 to 0.030 m³/m² (proximal portion) and from 0.016 to 0.080 m³/m² (distal portion). In unmyelinated group-IV nerve fibres, the values also showed a broad variation ranging from 0.001 to 0.011 m³/m² (proximal portion) and from 0.003 to 0.019 m³/m² (distal portion). The wide range of mitochondrial concentrations may reflect different energy consumption during receptive processes: nerve fibres with a low mechanical threshold and a high probability of excitatory events may be rich in mitochondria, whereas fibres with a high mechanical threshold and a low probability of excitatory events may be poor in mitochondria.     

Experimental evidence for interactions between bacterial peptidase and alkaline phosphatase activity in the Baltic Sea

From the observed pattern of aminopeptidase and alkaline phosphatase activities in the Baltic Sea, the question arose whether there is an interaction between the activities of both enzymes. In experiments with 0.8 m filtered seawater, the effects of commercial alkaline phosphatase on bacterial aminopeptidase, the effects of commercial peptidase on bacterial alkaline phosphatase activity (APA), and the effects of proteins, carbohydrates and inorganic nutrients on the activities of both enzymes were investigated.Addition of commercial alkaline phosphatase stimulated bacterial aminopeptidase activity and, similarly, the addition of commercial peptidase increased the APA in bacteria. The proteins, albumin and casein, stimulated aminopeptidase activity and APA simultaneously. Experiments using ammonium and glucose suggested that stimulation of APA by peptidase could be mediated by nitrogen and carbon availability. There were also some indications that stimulation of aminopeptidase activity by alkaline phosphatase functioned by catalysing phosphate release from organic phosphorus compounds.     

Anti-Auxin Enhance Rosa Hybrida L. Micropropagation

Shoots of rose cultivars Super Star and Sonia were multiplied for ten subcultures at 4-week intervals on solidified Murashige and Skoog's medium supplemented with 22.19 M benzylaminopurine + 1.07 M napthalene acetic acid + 0.05 M gibberellic acid. Addition of anti-auxins 2,3,5-triiodobenzoic acid (TIBA; 2.0 M) and 2,4,6-trichlorophenoxy-acetic acid (2,4,6-T; 0.39 M) into proliferation medium increased number of shoots per explant and length of shoots in both cultivars. Treatment with TIBA increased also number of leaves per shoot and leaf chlorophyll content.     

Kinetics of alkaline phosphatase activity and phosphorus availability for phytoplankton and bacterioplankton in lake plu\see (North German Eutrophic Lake)

Annual studies of kinetics of alkaline phosphatase (APA) activity and phosphorus availability for microplankton in the photic zone of an eutrophic lake are reported. The total APA activity of microplankton varied strongly. V_max was highest during summer P depletion, and in autumn and winter total APA activity was low. The total APA specific activity of the microplankton was also highest (average 3.55 pmole PO₄ ^3– ng ATP^–1 min^–1) when ambient orthophosphate concentrations were very low. Both V_max and specific APA activity were not dependent on the biomass of microplankton; they were strongly affected by P available for microplankton. A differential filtration technique was used for separation of microplankton into two size classes, i.e., algal, larger than 3m, and bacterial fraction with size 0.2–3.0m. The algal size fraction had lower specific APA activity (average 1.224 pmole PO₄ ^3– ng ATP^–1 min^–1) and higher K_M values (38.8mole × liter^–1) than microorganisms which were smaller than 3m (2.011 pmole PO₄ ^3– ng ATP^–1 min^–1 and 25.4mole liter^–1, respectively). The K_M values of free, dissolved APA (36.8mole liter^–1) indicated that free APA was probably released by algae. Phytoplankton were major APA activity producers in the photic zone of the lake from March to November, and their activity constituted, on the average, 48.6% of the total APA activity in the water. Bacteria were the dominant APA activity producers in winter (41.3–44.9%); however, during other periods they contributed significantly (average 21.7%) to total APA activity. When surplus constituted less than 10% of particulate P in seston, phytoplankton produced high specific APA activity, and when surplus P was higher than 15%, the specific APA activity of phytoplankton size fraction rapidly decreased. APA of the bacterial size fraction of the seston was not affected by P concentrations. Orthophosphate was a competitive inhibitor of APA produced by microorganisms of the size fraction larger than 3.0m, and increasing concentrations of inorganic phosphate caused an increase in K_M values. The hypothetical metabolic-coupling between phytoplankton and bacterioplankton in the phosphorus cycle in conjunction with carbon metabolism in the lake is discussed.     

Plantlet regeneration from callus initiated from flower buds in the wild species Allium senescens var. minor

Callus regeneration was observed from flower buds of Allium senescens var. minor inoculated in BDS, MS or B5 medium supplemented with 4.4 M benzyladenine alone or in combination with 4.5 M 2,4-dichlorophenoxy-acetic acid (2,4-d), with 2,4-d and kinetin (4.5 M/4.6 M) or with 5.3 M naphthaleneacetic acid. Ovules enlarged initially but the embryogenic tissue degenerated as callus development progressed from the nectar regions of the petals. Shoot buds and leaf primordia developed from the meristematic protuberances that originated from the surface of the callus. BDS medium with 4.5 M 2,4-d and 13.3 M BA was most suitable for shoot multiplication. The regenerated shoots were rooted in respective liquid medium without any growth regulators and successfully transferred to soil with 90% survival rate.Abbreviations BA N⁶-benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid     

Mortality of marine bacterial strains in seawater

As an approach for assessing the dynamics of bacterial population in seawater, the survival of five isolated marine bacteria strains was assessed by the disappearance of radioactivity in the cold trichloroacetic acid (TCA)-insoluble fraction from a previously³H-labeled culture. Metabolic activity during survival experiments was assessed by the measurement of electron transport system (ETS) activity. Fractionated filtration was used to assess the grazing mortality. The particulate fraction that passed 2.0 m and was retained in 0.2 m was the main cause of mortality.     

Storage,mobilization and interrelations of starch,sugars, protein and fat in the ray storage tissue of poplar trees

The seasonal pattern in starch, various sugars, protein, and fat, and their interrelationship, has been followed in 3-year-old branch wood of poplar trees (Populus x canadensis Moench robusta) under natural site conditions. The deposition of starch, protein and fat proceeds at different times. Starch accumulates from May until October, fat mainly during the summer months, and protein when the leaves are yellowing in September and October. The maximum concentrations in the branch wood were 15–18 g starch, 6–9 g protein, 4–8 g fat, 10–15 g sucrose, and up to 30 g total sugars per milligram dry weight (DW). During starch deposition periods no increased sucrose level is found in the tissue. The maximum daily starch deposition rate was 0.2–0.4 g starch/day/mg DW of wood. During starch hydrolysis in late autumn and winter, a dramatic increase in sucrose and its galactosides is measured (up to 15–27 g/mg DW in total). In early spring, before budbreak, the concentrations of these sugars diminishes sharply. In contrast to this clear-cut starch-to-sugar conversion in autumn no significant starch-to-fat conversion is detected. An elevated content of free glycerol, however, is found in winter. In spring, starch and storage protein are mobilized completely, or almost completely, in poplar twig wood. A noteworthy pool of maltose is found transiently during autumn (up to 8 g/mg DW) and again in spring. The results demonstrate that the individual storage materials, e.g. starch, protein, and fat, are accumulated fairly independently in the wood storage parenchyma. Tissue sugar levels, in contrast, appear to be closely related to the seasonal variations in starch content, on the one hand, and to the acclimation and deacclimation of the cells, on the other. The interrelations of the storage materials and sugars are discussed.     

Purine and pyrimidine base and nucleoside concentrations in human cerebrospinal fluid and plasma

Purine and pyrimidine base and nucleoside levels were determined in adult human lumbar (CSF) and plasma by reversed-phase high performance liquid chromatography (HPLC). Guanine, thymine, cytosine and uracil were not detectable (<0.1 M) in human CSF or plasma. Adenine was detectable in plasma (0.3 M) but was not found in CSF (<0.2 M). Hypoxanthine and xanthine levels in CSF were each approximately 2.5 M. Plasma levels of hypoxanthine and xanthine were considerably lower (0.4–0.6 M). Purine and pyrimidine ribouncleosides in human CSF were less than or equal to 0.2 M with the exception of uridine which was present at concentrations of 2–3 M. Although low concentrations of thymidine and deoxyuridine (0.2 M) were present in human plasma, purine and pyrimidine deoxyribonucleosides were less than 0.1 M in human lumbar CSF.     

The in vitro culture of Gerbera aurantiaca

Multiple shoot cultures were obtained from elongating axillary buds of Gerbera aurantiaca Sch. on the medium of Murashige & Skoog (MS) supplemented with 5 M benzyladenine. Shoots rooted in vitro on MS medium supplemented with 5 to 10 M indolebutyric acid or 5 to 10 M -naphthaleneacetic acid. Plantlets were hardened in a mist bed and transferred to soil.     

Biosynthesis of glycoproteins in Candida albicans: Biochemical characterization of dolichol phosphate glucose synthase

A mixed membrane fraction isolated from C. albicans yeast cells catalyzed the transfer of glucose from UDP-Glc into three classes of endogenous acceptors: glucolipid, glycoprotein and lipid-linked oligosaccharides. About 80 of the total radioactivity transferred into these products corresponded to the glucolipid which was identified as dolichol phosphate glucose by several criteria. The remainder was detected in about equal proportions in the other two fractions. Conditions that stimulated or inhibited glucolipid synthesis did not affect the extent of glycoprotein labeling. The synthesis of dolichol phosphate glucose exhibited a K_mof 104 M UDP-Glc and was stimulated by Mg²⁺but not by Mn²⁺or Ca²⁺. The latter cations were, however, better stimulators of glycoprotein labeling than Mg²⁺. Most nucleotides strongly inhibited the synthesis of dolichol phosphate glucose, UMP being a competitive inhibitor with a K_iof 100 M. The dolichol phosphate glucose synthase reaction was reversed about 57 by 0.62 mM UDP but not by UMP.     

Minimal growth in vitro conservation of coffee (Coffea spp.)

We have studied the influence of low concentrations of 6-benzyladenine on growth limitation, in order to preserve coffee germplasm through a microcutting collection. Concentrations of 0 M, 1.3 M and 4.4 M were compared in four species: Coffea congensis, C. canephora, C. liberica and C. racemosa. After six months, microcutting behaviour varied between the different treatments, and a species effect was observed. The slow growing species (C. liberica and C. congensis) needed 1.3 M; the others coffee species (C. canephora and C. racemosa) exhibited moderate caulogenesis on 6-benzyladenine-free medium. Zero and low concentrations did not affect survival rates. In conclusion 1.3 M seems most appropriate for conserving all four species.Abbreviation BA 6-benzyladenine     

Trois nouveaux Nadejdolepis Spasskii & Sasskaya, 1954 (Cestoda: Hymenolepididae) parasites de Charadrii (Aves) de Tasmanie

Three species of Nadejdolepis from Tasmania, Australia, are described and illustrated. N. burgessi n. sp., a parasite of Charadrius ruficapillus, is 4-6 mm long, with rostellar nitiduloid hooks 63-66 m long, a short evaginated cirrus 13-16 m long with a short collar of thin spines 1 m long, a narrow and tubular sclerotinoid vagina 40-50 long and 3-4 m in diameter with a little ampulla 3-5 m in diameter at the proximal end, and a membranous atrial segment with smooth, short (1 m) and compact spines which are sometimes difficult to observe. N. smithi n. sp., a parasite of 40-50 long and 3-4 m in diameter with a little ampulla 3-5 m in diameter at the proximal end, and a membranous atrial segment with smooth, short (1 m) and compact spines which are     

Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids

Leaf explants of Paphiopedilum phiIippinense hybrids (hybrid PH59 and PH60) directly formed adventitious shoots from wound regions within 1 month, when cultured on modified Murashige and Skoog medium (1/2-strength macro- and full-strength micro-elements) free of plant growth regulator in darkness. The combinations of 2,4-dichlorophenoxyacetic acid ((2,4-D) acid (0, 4.52 and 45.25 M) and 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ) (0, 0.45, 4.54 and 22.71 M) were used to test their effects on direct shoot bud formation from two types of explants (1.5-cm long intact leaf explants and 0.5-cm long leaf segment explants). In hybrid PH59, 4.54 M TDZ increased mean numbers of shoots per explant with leaf segment explants. In hybrid PH60, 4.52 M 2,4-D plus 0.45 M TDZ promoted direct shoot bud formation from leaf segment explants. In addition, three treatments (4.52 M 2,4-D, 22.71 M TDZ, 4.52 M 2,4-D plus 4.54 M TDZ) gave a higher response than control on mean numbers of shoots per explant with intact leaf explants. Healthy plantlets each with one to three roots were obtained from leaf-derived shoots after transfer onto a hormone-free medium for 22 months. These plantlets were acclimatized in a greenhouse and grew well with 100% survival rate.     

Der praecorneale Glykosaminoglykan-Film beim Haushuhn

Summary The anterior cornea epithelium of the house-hen consists of 5 layers. The outermost layer has a villus border. The 0.2 long microvilli are 0.04–0.05 thick and are not more than 0.1 apart. A film of ruthenium red stainable glycosamine Glycans of 0.26–0.3 thickness lies on the free surface. The microvilli protrude into this film. This film is responsible for the reflecting shine of the cornea. The tear secretion can be kept at a low level as a part of the fluid is bound to the acid mucopolysaccharides. The molecules for this film are probably synthesized in the outermost cell layer.

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Der praecorneale Glykosaminoglykan-Film beim Haushuhn

Zusammenfassung Das vordere Corneaepithel des Haushuhnes besteht aus 5 Schichten. Die oberflächlichste Lage hat einen Zottenbesatz. Die 0,2 langen Mikrovilli haben eine Dicke von 0,04–0,05 und einen Abstand von höchstens 0,1 voneinander. Auf der freien Oberfläche liegt ein mit Rutheniumrot anfärbbarer Film von Glykosaminoglykanen von 0,26–0,3 Dicke. Die Mikrovilli ragen in diesen Film hinein. Dieser Film ist für den spiegelnden Glanz der Cornea verantwortlich. Die Tränensekretion kann sehr gering sein, weil ein Teil der Flüssigkeit an die sauren Mukopolysaccharide gebunden ist. Die Moleküle dieses Films werden wahrscheinlich in der oberen Zellage synthetisiert.

     

A decrease in intracellular zinc level precedes the detection of early indicators of apoptosis in HL-60 cells

Low extracellular zinc concentrations have been associated with the induction of apoptosis. To assess the relationship between intracellular zinc concentration and the rate of apoptosis, cells were grown in media containing 0.5, 25, or 50 M zinc and analyzed by flow cytometry or fluorescence microscopy. Cells grown in 0.5 M zinc medium over 48 h showed a successive decrease in intracellular zinc concentration measured by the zinc-specific fluorophore, zinquin. After 18 h in 0.5 M zinc medium, rhodamine 123 retention decreased. However, the addition of 10 M zinc to the 0.5 M medium before 16 h in culture restored rhodamine retention in the cells. After 30 h there was an increase in the number of cells cultured in 0.5 M zinc medium that bound annexin V-FITC. These data indicated that decreased intracellular zinc concentration preceded early markers of apoptosis, with alterations in mitochondrial transmembrane potential preceding the loss of polarity in the cell membrane.     

A between-river comparison of extracellular-enzyme activity

River-water extracellular-enzyme activity in the lowland Rivers Ouse and Derwent, northeast England, had much in common. In both rivers, the mean enzyme activities over 15 months differed in the following order: leucine aminopeptidase > phosphatase > -D-glucosidase > -D-galactosi-idase and -D-xylosidase. None of the five enzymes assayed had significant between-river difference in activity, and there was significant between-river correlation of -D-glucosidase, phosphatase, and leucine-aminopeptidase activity. The common enzyme regimes were probably more due to between-river similarity of planktonic microbiota than to similar physico-chemical conditions. The potential for glucose uptake by bacterioplankton closely followed -D-glucosidase activity in magnitude and periodicity. The potential for leucine uptake, however, was much less than leucine-aminopeptidase activity; hence rate of leucine release probably did not limit leucine uptake. There was an appreciable and highly variable proportion of free (<0.2 m) enzyme activity in river water; ranges were -D-glucosidase 10–30%, phosphatase 53% to apparently 104%, and leucine aminopeptidase 22–98%. These free enzymes did not necessarily originate from planktonic microbiota and may explain the fairly loose coupling between whole-water enzyme activity and microbial variables. Marked downstream increase in enzyme activity, along about 104 km of the River Derwent, was found on only one of three sampling days; hence the single site used for regular sampling was reasonably representative of most of the river.     

In vitro shoot regeneration from leaf explants of Roman Chamomile

Murashige and Skoog (1962) medium supplemented with 1.0 to 4.5 M of BA and 1.0 M of NAA induced adventitious bud formation and shoot development in leaf explants of Roman Chamomile. A higher number of adventitious buds was observed at the proximal end of the explants. Plantlets were replicated and multiplied on MS medium supplemented with 2.25 M of BA and 0.6 M of IAA. Plantlets were rooted on MS medium supplemented with 0.5 M of IBA and successfully weaned in vivo. The plants grew to maturity with high uniformity and no morphological signs of somaclonal variation.